Rapid Micro Methods News

Our news pages will keep you informed of press releases and news articles on RMM technologies, updates from technology suppliers, reviews of recent publications and presentations, and what's changing in the world of rapid methods. You can also follow our news posts on Twitter, Facebook, LinkedIn and RSS.

Tuesday, July 28, 2015

DigiPath Labs, the cannabis testing subsidiary of DigiPath, Inc., is combining efforts with True North Laboratory of Ann Arbor, Michigan, and Romer Labs, a leading global supplier of diagnostic solutions for food-borne pathogens, to investigate the applicability of utilizing Romer Labs' rapid diagnostic test kits to screen for food-borne pathogens and quantify mycotoxins present in cannabis.

"This joint effort is expected to result in the first validated kit-based assays for food-borne pathogen and mycotoxin quality assurance testing on cannabis," says Dr. Cindy Orser, DigiPath Labs' Chief Science Officer. "As more states approve medical marijuana, the importance of validated studies such as this one planned for quality assurance will become of keen interest to officials from state public health departments who are responsible for ensuring the safety of medical marijuana."

During the one-week study, Dr. Orser of DigiPath Labs and two scientists from Romer Labs will execute the validation studies at True North Laboratory in Ann Arbor, Michigan, where access to marijuana provides the plant matrix to conduct the testing to validate Romer Labs' AgraQuant® Ochratoxin A ELISA System, which is quantitative from 2 to 40 ppb, and its AgraStrip® Aflatoxin Lateral Flow System, which features a 20 ppb aflatoxin cutoff sensitivity, and the fully quantitative AgraStrip® Aflatoxin WATEX test, which utilizes a water-based extraction process. All solutions are capable of quantifying mycotoxins within specifications of Nevada's cannabis quality assurance testing guidelines.

Friday, July 24, 2015

The BBI Group (BBI) announced it has developed in collaboration with the Defence Science and Technology Laboratory (Dstl) and licensed, through technology transfer company Ploughshare Innovations, a rapid test for Ebola. The assay, which is in the process of being verified to allow EUA submission, will be used to test patients suspected of Ebola infection to screen and potentially help with how they are subsequently managed. The test, utilising lateral flow technology, can provide a result in approximately 20 minutes.

More than one year after the initial outbreak, the number of Ebola cases sits at over 27,000 cases reported and 11,000 deaths in West Africa to date (1). Accurate diagnosis currently relies on skilled laboratory staff and equipment, largely unavailable in the field, resulting in delays in diagnosis and difficulty containing spread of the virus (2).

Using lateral flow technology, BBI’s assay will make testing simpler and quicker than assays such as RT-PCR that requires technical infrastructure. With minimal training, local health teams can complete the test at bedside and have a result in 20 minutes. In field data suggests the test could be used as a negative agreement ‘rule-out’ test and screening method (3). In the future, such a test could possibly be deployed at borders to monitor and control spread of the virus.

Through collaboration with partners, a lateral flow test has already been evaluated in the field. In trials in Sierra Leone, of 138 participants enrolled in testing, a 100% detection rate was achieved. At a low positive the test achieved 100% sensitivity and 92% specificity, and at a high positive, 100% sensitivity and 97% specificity (3), making the BBI assay potentially more sensitive and specific than other lateral flow or dipstick type assays currently available to date.

Using the unique integrated AtomoRapid™ platform, the number of components in the kit is reduced, simplifying the process for end users whilst reducing the risk of user error and cross contamination.

Aiming for a 3rd quarter launch following regulatory approval, the test will be available for immediate deployment in affected areas. BBI will then roll out to all global territories following their respective regulatory registration and acceptance.

In addition, a second version of the test will be evaluated in combination with the Novarum DX Smartphone reader. Allowing users to read and share test data using smartphone technology. This development could potentially allow instant connectivity of patients and field workers with medical experts, while also utilising built in GPS capability to track and monitor disease ‘hot spots’.

Leigh Thomas, Chief Commercial Officer commented “BBI has developed high performance lateral flow tests for some of the world’s leading diagnostic companies for over 25 years. Our expertise and flexibility has allowed us to quickly deploy a team and develop a rapid test in 9 months which, based on field testing, promises to have an immediate and positive impact on the Ebola crisis. We are pleased that our test will meet an immediate need in the field and ultimately improve the lives of others”.

NHS Fife has selected Fast-track diagnostics Respiratory 21 assay for the identification of respiratory viruses in its laboratories. The assay will assist NHS Fife in delivering fast and accurate diagnoses to hundreds of patients.

Fast-track diagnostics develops and manufactures more than 60 different assays in 32 and 64 reaction kit sizes, detecting more than 150 different pathogens. With its multiplexed approach, Fast-track diagnostics kits enable the simple, rapid detection of most of the clinically relevant pathogens present in a sample. The kits are a cost-effective way of delivering a comprehensive diagnostic service and streamlining laboratory workflow and are compatible with a wide range of open-system thermo cyclers.

In the comparison study in Fife, a set of 68 samples were tested using the Fast-track diagnostics Respiratory 21 assay and a competitor assay. Both multiplex real-time PCR kits detected the same wide range of viruses, and the Fast-track diagnostics assay detected a positive Rhinovirus and Adenovirus missed by the competitor assay. In addition the Fast-track diagnostics multiplex includes the detection of Mycosplasma and Parechovirus and so was selected for routine use. Furthermore, the shorter assay time (2.5 hours from sample to result), made it possible to perform two runs per day with a high level of accuracy and reliability.

“NHS Fife is continually looking to improve upon existing diagnostic methods to offer the best possible service to clinicians and patients,” said Bob Jones, BMS Professional Manager at NHS Fife. “The shorter assay times streamline our workflow and the increased number of pathogens detected allows for greater accuracy and clinical relevance. Overall the Respiratory 21 assay from Fast-track diagnostics showed considerable advantages enabling more effective patient care and treatment”.

Verax Biomedical announced it has renewed its exclusive worldwide commercialization agreement with Fresenius Kabi USA focused on improving blood collection, separation, safety and availability. The agreement is to market, sell and distribute Verax's unique PanGenera® Detection (PGD) test - a rapid in vitro diagnostic test used to detect bacterial contaminants in donated platelets. The Verax Platelet PGD® test is the only rapid test on the market cleared by the FDA for all commonly available U.S. platelet types.

"Verax Platelet PGD® test is the industry standard when it comes to the detection of bacteria in platelets, and we are pleased to continue our partnership with Fresenius Kabi to combat the severe risk of bacterial contamination in the U.S. blood supply - the number one infectious risk to platelet transfusion recipients," said Verax Biomedical CEO Jim Lousararian. "Fresenius Kabi is a leader in the development of products that improve blood collection and safety worldwide, and we look forward to continuing our work with them to improve health care outcomes by reducing transfusion reactions associated with bacterial contamination."

"The Verax Platelet PGD® test is an essential safety tool that every hospital should use before transfusions," said Dean Gregory, president, Medical Devices, Fresenius Kabi USA. "This agreement reaffirms our commitment to Verax technology, and is yet another example of how we are supporting improved safety in transfusion medicine."

The Verax PGD® test, based on proprietary technology developed by Verax, consists of an easy-to-use disposable test device and reagents that work together to detect the presence of bacterial contaminants in platelets. The test can be performed in less than 30 minutes and is designed for use in hospitals, cancer centers and other sites of care as a safeguard prior to transfusion.

Annually, more than 6 million platelet doses are transfused worldwide, and approximately 1 in 2,000 doses are contaminated by bacteria. The presence of bacteria in platelets can pose serious risks for patients, including sepsis—a life-threatening infection in the blood stream. The test is a cost effective approach to platelet bacterial safety.

Produce growers and shippers who say test-and-hold screening takes too much time may not have that excuse much longer as a new method promises to reduce two-day sample preparation time to two hours.

The new method earned a group of researchers at Purdue University the grand prize in the Food and Drug Administration’s first Food Safety Challenge contest. The agency awarded the Purdue group $300,000 in what FDA officials have said will be an annual competition.

Pronucleotein Inc., a San Antonio biotechnology company, won the runner-up title and a $100,000 prize for a portable rapid pathogen screening system that researchers say will provide test results in about 30 minutes.

The researchers from Purdue, led Michael Ladisch, have been working on their salmonella microfiltration concentration preparation method for about seven years, according the FDA’s contest blog.

For the blog, FDA staffers asked a representative of each of five finalist teams to answer five questions, including what insights the researchers had gleaned from the competition.

Both Purdue’s Ladisch and John Bruno, vice president and chief technology officer at Pronucleotein, said the contest helped them better understand the challenges food safety officials face.

“The biggest insights have been on the needs of (FDA) with respect to the large number of samples that must be processed, the importance of rapid detection of pathogens in fruits and vegetables, and detection sensitivity required in FDA laboratories,” Ladisch said in the blog.

Bruno said the competition gave him and research partner James Byron, CEO of Ponte Vedra Beach, Fla.-based medical laboratory Xgenex, a better idea of how to “ease the burdens of FDA inspectors at ports.”

“We now have a much better understanding of the problems the FDA faces in attempting to prevent foodborne illnesses from imported foods and we are better able to tailor our solution for FDA screening scenarios,” Bruno said in the blog Q&A.

The other three researchers and projects selected as finalists, which each earned $20,000 in the initial round of the competition, were:

University of Illinois and Purdue — a group of researchers are developing a “point-of-care” handheld system that incorporates sample preparation and pathogen detection. The team estimates that concentrating the cells takes about two hours and detection adds another 45 minutes.

Bart Weimer, of the University of California-Davis and Mars Inc., is perfecting a means of capturing and concentrating salmonella using a glass bead. The process takes about three hours.

Auburn University — Brian Chin and others developed a biosensor that can detect pathogens directly on the surface of foods in 2-12 minutes.

Micro Imaging Technology, Inc. announced that, through its collaboration with Northern Michigan University (NMU), it has developed a testing protocol that allows for detection of pathogen species from liquid cultures. Dr. Josh S. Sharp, Ph.D., assistant professor at the NMU Department of Biology in Marquette, Michigan, has been spearheading the collaboration between MIT and NMU since October of 2013. He is researching clinical applications of the MIT 1000 System, particularly the pathogens Staphylococcus aureus (S. aureus) and Methicillin Resistant S. aureus (MRSA).

"Many of the current agar plate based identification methods require 16-24 hours of growth before identifications can be made," said Dr. Sharp. "Our work with MIT and its rapid laser-based identification system, the MIT 1000, has effectively eliminated the need for that time-consuming step and reduced the time required for pathogen sample preparation and identification to 4-6 hours given a sufficient starting inoculum. This new protocol allows for detection of Staphylococcus species from liquid cultures and has provided important groundwork to facilitate rapid enrichment of target bacteria for identification using the MIT 1000."

"The MIT 1000 has already proven to be one of, if not the fastest and least expensive method for pathogen testing out there," said Jeff Nunez, President of MIT. "This recent innovation in capturing target bacteria, eliminating the need for agar plate culturing, should prove to be a highly significant leap forward for the MIT 1000 in both clinical and food safety applications."

Friday, July 17, 2015

Just four companies worldwide are cleared to trial their quick tests for the deadly Ebola virus.

And Crystal Diagnostics hopes to soon join its Broomfield neighbor Corgenix Medical Corp. on that short list.

When Corgenix's ReEBOV product became the first rapid Ebola test to receive the World Health Organization's blessing this year, it was among fewer than 20 Ebola-related products submitted for the agency's assessment.

Now Crystal Diagnostics, which developed a liquid crystal-based commercial product to test for pathogens in food, is adapting its device to test for diseases such as the Ebola virus.

After a couple months of testing surrogate products that mimicked the Ebola infection, Crystal Diagnostics' test was successful in detecting a live-virus sample in less than 20 minutes — putting it on par with the Corgenix test, officials said.

"It's very fast, very accurate, and our system is very automated," said Crystal Diagnostics CEO Jim Bruce, whose company is less than a mile from Corgenix.

And where Corgenix fills a need in providing an inexpensive test that can be used in remote locations, Bruce believes Crystal Diagnostics can create an automated product better suited for hospitals, airports and clinics. Crystal Diagnostics' test would cost $30, or about double the price of ReEBOV, Bruce estimated.

But to get to that point, Crystal Diagnostics will need to conduct additional research, including field testing in Africa, where the virus remains active in Guinea, Libera and Sierra Leone. The company has submitted a grant application to the Paul G. Allen Family Foundation, which helped fund development of Cogenix's quick test. It also plans to apply to the Biomedical Advanced Research and Development Authority.

If all goes well on the research front, Crystal Diagnostics could receive approvals from WHO and the Food and Drug Administration in less than a year, Bruce said.

Through this process, Bruce and Crystal Diagnostics' officials have knocked on their neighbor's door, querying Corgenix about its path to approval. Corgenix's test can detect the virus in less than 15 minutes, instead of hours or days.

Former Corgenix CEO Doug Simpson, who now oversees the company's infectious-diseases group, is taking a many-hands-make-light-work approach when it comes to Ebola-related technologies.

"It's not just a Corgenix technology," Simpson said, noting ReEBOV was developed in concert with academic entities, including the Viral Hemorrhagic Fever Consortium. "If there are other ways that (VHFC reagent) could be of use, maybe in different segments of the market, that's fine."

Thursday, July 16, 2015

Alere Inc., a global leader in rapid diagnostic tests, today announced that the U.S. Food and Drug Administration (FDA) has granted CLIA waiver for the Alere™ i Strep A test. The test, which was cleared for marketing by the FDA in April 2015, is the first molecular platform that detects Group A Streptococcus (GAS) bacteria in 8 minutes or less.

With CLIA waiver, the Alere i Strep A test will be available in a broad range of healthcare settings, including physician offices, hospital emergency rooms, and retail and walk-in clinics in the United States.

"Strep A is highly transmissible, and prompt treatment is critical to reducing the spread of the infection to others. With CLIA waiver for the Alere i Strep A test, we can help link more people to early treatment by bringing rapid molecular diagnosis to even more settings where they seek health care," said Avi Pelossof, Global President of Infectious Disease at Alere.

The Strep A test is the second assay on the Alere i molecular platform, which initially received 510(k) clearance for the detection and differentiation of influenza A and B virus in 2014. In January 2015, Alere i Influenza A & B became the first-ever molecular test to receive CLIA waiver.

About the Alere i Strep A testAlere i Strep A detects Group A Streptococcus (GAS) bacteria in throat swab specimens using Alere's proprietary Molecular In Minutes™ (MIM) isothermal nucleic acid amplification technology (iNAT). Unlike polymerase chain reaction (PCR) tests, iNAT does not require lengthy and complex thermocycling or DNA purification and can therefore deliver PCR-caliber results more quickly – and in a broad range of settings.

Other assays currently in development on the Alere i platform include respiratory syncytial virus (RSV), C. difficile and Chlamydia/Gonorrhoea.

About Strep AGroup A Streptococcus (Group A Strep, or GAS) bacteria spread through contact with droplets from an infected person's cough or sneeze, and live in a person's nose and throat. Most GAS infections cause relatively mild (noninvasive) illnesses such as strep throat, scarlet fever, and impetigo (a skin infection). More than 10 million non-invasive GAS infections (primarily throat and superficial skin infections) occur annually in the U.S.[1] Occasionally, these bacteria can cause severe and even life-threatening (invasive) diseases. Cases of invasive GAS infections, such as necrotizing fasciitis and streptococcal toxic shock syndrome, occur less frequently but are associated with higher rates of deaths.[1]

Researchers at the New York State Department of Health's Wadsworth Center have developed a multiplex real-time PCR assay to detect the enterotoxin genes of various Staphylococcus aureus strains and enable rapid outbreak detection and control.

The assay, along with a singleplex positive extraction control to detect inhibition, was published online in Molecular and Cellular Probes.

The protocol also includes a modification of the MagNA Pure Compact automated extraction method from Roche that allows improved S. aureus detection from stool samples.

Staphylococcal contamination caused 570 outbreaks of food poisoning between 1998 and 2010, and Staphlococcus aureus type B is considered a potential bioterrorism agent. Wadsworth Laboratory is New YorkState's reference lab tasked with responding to these types of public health threats.

"It's important for us to be able to quickly determine the cause of an outbreak of illness so that it doesn't spread throughout the public," Christina Egan, corresponding author on the study and director of the Wadsworth Center Biodefense Laboratory, told GenomeWeb in an interview.

"Our previous methods were based on culturing the agent from different food sources — which could take days — and we really wanted to be able to more rapidly identify a causative agent," she said.

In the literature there are a few assays for S. aureus in food, but Egan believes this is the first that combines multiplex real-time PCR with the automated extraction process needed to process larger numbers of samples.

As described, the assay focuses on clinical patient stool samples, but Egan said her group has also validated it in several different types of food matrices.

The assay detects staphylococcal enterotoxins A through E as well as toxic shock syndrome toxin production genes.

"That's really key because we are testing directly from patient specimens and foods and we don't have an incubation step, so we really want to be able to have as efficient a reaction as possible to detect any nucleic acid that's present and has a toxin sequence," Egan said.

The researchers tested the assay on 47 strains of microorganism, and found 100 percent specificity and a sensitivity of 8 to 20 colony forming units per real time PCR in clinical stool matrix.

The researchers are not planning to pursue patent protection for this particular assay, Egan said.

"We've published it and made it available so that other public health labs and clinical laboratories can utilize it to improve their diagnostic capability," she said.

However, she noted that the lab is glad to work with companies that are developing diagnostics in this area. "We've done that in the past and that's something that we're always willing to do."

S. aureus is not represented in the multiplex GI panels that have recently become commercially available, possibly because it is so self-limited, but Egan noted that "it would be great to develop a panel that includes staph enterotoxin."

Wadsworth houses several regulatory programs, including one that grants permits to both environmental and clinical laboratories, including CLIA certification. It is also involved in development of molecular diagnostics, particularly ones aimed at "testing more agents with less specimen," Egan said.

For example, Wadsworth has collaborated with Columbia University and Northrup Grumman to develop a biothreat agent diagnostic using mass spectrometry. With Akonni Biosystems, Wadsworth has worked on diagnostic viral arrays, including ones for meningitis/encephalitis and influenza, as well as a tuberculosis assay, as previously reported.

Mass food poisoning and bioterror

The Molecular and Cellular Probes study also notes that the S. aureus assay was used to detect an outbreak in New YorkState.

In that case, attendees of a large event at Chuang Yen Buddhist Monastery in Putnam County consumed tainted food.

After eating at the event, one large contingent from the New York City area then boarded several buses for a visit to the Monroe-Woodbury outlet mall.

"By the time they got to the mall, many of them were really ill," Egan said. The presence of such a large number of sick people, some of whom could hardly walk according to news coverage from the time, triggered an alert and the county health department was then able to retrieve food from the event. All told, about 150 people became ill, according to another report.

"What was unfortunate was that many of the [food vendors] had traveled from the tri-state area and we think that the food was stored under conditions that allowed Staph enterotoxins to be produced," Egan said.

Deliberate mass food poisoning is also a biothreat that Wadsworth vigilantly monitors, and Staph enterotoxin type B is on the US Centers for Disease Control and Prevention's Bioterrorism agents list.

S. aureus enterotoxin illness is self-limiting and usually non-fatal, but it could potentially be used to disrupt the food supply, a situation which one study estimates could cost $30 billion to remedy.

Although there are a number of known non-terrorist biocrime cases, there have only been two confirmed incidences of terrorist use of biological weapons since 1945.

Egan mentioned that one was the 1984 case of mass food poisoning in Oregon perpetrated by a cult, referred to as the Rajneeshee bioterror attack.

In that incident, salmonella was cultured and then used to contaminate food and condiments at 10 restaurant salad bars. The attack ultimately sickened 751 people, but failed to exert the desired political effect of disrupting a local election.

The independent laboratory study testing was conducted by Marshfield Food Safety as part of the AOAC-RI independent third-party evaluation and validation processes. "With recent events involving Listeria outbreaks we believe that environmental testing plays an important role for food safety," said Mary Duseau, Roka chief commercial officer.

Roka Bioscience is a molecular diagnostics company focused on developing and commercializing advanced testing solutions for the food safety testing market. Our Atlas Detection Assays incorporate our advanced molecular technologies and are performed on our "sample-in, result-out" Atlas System that automates all aspects of molecular diagnostic testing on a single, integrated platform. The Atlas System and Detection Assays are designed to provide our customers with accurate and rapid test results with reduced labor costs and improved laboratory efficiencies.

"The AOAC approval further validates our test as an invaluable tool to food producers," said Ed Bradley, Neogen's vice president for Food Safety. "ANSR is the fastest DNA-definitive pathogen assay available — with results in only 10 minutes. Compared to the three hours other methods such as polymerase chain reaction, or PCR, take to produce DNA-level results, that's a huge difference in a laboratory's workflow, and the operations of a food producer as a whole."

Combined with ANSR's single-step enrichment, Neogen's new pathogen detection method for L. monocytogenes can provide definitive results in as little as 17 hours for environmental samples from the time the sample is taken. The new test also utilizes Neogen's LESS Plus Medium, which can be autoclaved — allowing for larger batches of media to be prepared prior to use.

The approval covers the use of the ANSR system to detect L. monocytogenes in the following sample types: hot dogs, Mexican-style cheese, cantaloupe, guacamole, pasteurized liquid egg, sprout irrigation water, and sponge samples from stainless steel surfaces.

Neogen's line of ANSR products also includes AOAC Research Institute-validated ANSR for Listeria and ANSR for Salmonella.

Thursday, July 9, 2015

Crystal Diagnostics (CDx) has successfully detected live Ebola virus with its unique liquid crystal-based rapid pathogen detection system. Those tests were conducted at the University of Texas Galveston Medical Branch Biosafety Level 4 (BSL-4) facilities. Tests demonstrated detection at variable concentrations of live Ebola Zaire in serum samples in less than 20 minutes.

CDx sells rapid pathogen systems in the Food Safety market to quickly and accurately detect food borne pathogens in a variety of products including meat and produce. The company's systems are in use in commercial test labs, government agencies, universities, and research facilities, as well as food processors. The Ebola detection project is the first application outside of the food safety arena for CDx.

According to Robert F Garry, PhD, Professor of Microbiology and Immunology at the Tulane University School of Medicine, and Principal Investigator of the Viral Hemorrhagic Fever Consortium (VHFC), "The CDx system promises advantages over other rapid Ebola detection systems, in terms of automated reader results, speed, and consistency."

President and CEO of Crystal Diagnostics, Jim Bruce, stated that, "We are delighted with the initial results, and following field trials and US Food and Drug Administration device approval, we look forward to contributing to the fight against this deadly disease."

CDx has applied for grants from the Paul Allen Foundation and the Biomedical Advanced Research and Development Authority (BARDA) division in the U.S. Department of Health and Human Services to complete equipment automation and development of the Ebola test.

A team of researchers in Salt Lake City have conducted a health economics analysis on the effectiveness of using the BioFire FilmArray Respiratory Panel for diagnosing influenza in children who show up in the emergency room.

Published last month in The Pediatric Infectious Disease Journal, the study employed a decision analytic model to compare costs and outcomes of flu diagnosis using traditional PCR, direct-fluorescence antibody, rapid antigen tests, and the FilmArray RP.

Costs and benefits were found to hinge on the price and accuracy of the tests and the prevalence of flu, and were somewhat different for children younger than 3 compared to those between 3 and 18.

Ultimately, however, rapid multiplex PCR was determined to be the most effective strategy, the study concluded.

While it is associated with significant cost, the authors suggested that the cost is below a "willingness to pay" threshold for accepting new technologies that improve patient quality of life.

Health economics modeling for pediatrics is somewhat unique, Richard Nelson, a health economist and first author on the study, told GenomeWeb in an interview this week.

Typically a model will consider costs to three agents — the patient, the provider, and the payor.

In pediatrics, however, the group "patient" doesn't quite capture the dynamics, as the patient is sometimes non-verbal and is not usually the decision maker.

"There can be a burden or cost to caregivers that is separate to the cost to patients in this case," Nelson, who is affiliated with the Salt Lake City VA Medical Center and the University of Utah School of Medicine, explained.

This can be the case at the other end of the age spectrum as well, in geriatric patients. When a patient can't talk, she can't describe her symptoms, so molecular diagnostics may be particularly useful.

"A lot of the patients that get tested with our technology are in the non-verbal stages of their lives, so doctors are much more comfortable using a syndromic approach," he said, noting that children's hospitals have been the early adopters of the firm's technology.

Nelson said the modeling study was initiated in response to requests for data from clinicians at Primary Children's Hospital in Salt Lake City, which is the pediatric teaching hospital for the University of Utah School of Medicine.

"We had recently introduced [the FilmArray] here … so we wanted to compare the potentially increased benefit with the potentially increased cost," he explained.

Among the many authors on the study were experts in diagnostics, pediatric infectious disease, emergency department providers, and pathologists, he said, so it was very much "team-based science."

A few had some connections with BioFire in the early days, with one holding intellectual property and receiving royalties from the company. Nelson said this helped the study because these authors had special insights into the technology, but he asserted the study was completely independent.

The analysis was limited to the potential costs and benefits of detecting influenza, since there is not a large amount of data yet available on other targets, Nelson said. Expanding the model to include those other targets may enhance the estimated cost-effectiveness.

The results can also perhaps be expanded to other rapid diagnostics that may have similar accuracy, he said, as well as ones that potentially have a lower cost. These could include influenza-specific rapid molecular diagnostics like Alere i or Roche Liat.

Psychological benefits

Both Nelson and Stevenson pointed out what appears to be a growing realization in the field — the synergy between disease identification and patient satisfaction.

And in pediatrics, this force may be particularly pronounced, Nelson noted.

"One of the benefits of more accurate information is that it has the potential to alleviate anxiety or fear," he said.

For example, if new parents bring a baby to the emergency room with a high fever and cough, knowing that the baby has respiratory syncytial virus versus parainfluenza may not change the treatment course, but a specific diagnosis could provide the parents more comfort than saying, "it's probably a virus."

BioFire's Stevenson pointed out that patient satisfaction metrics may soon become part of healthcare evaluations though the Affordable Care Act.

This zeitgeist was also alluded to by BioMérieux's Christine Ginnocchio during a recent conversation with GenomeWeb, but Stevenson said it is not officially part of the BioFire messaging.

However, given the assumption that "the psychological value of the test is huge," the firm is now sponsoring studies to measure how its products affect patient satisfaction.

Stevenson said he anticipates increasing that message in marketing efforts in the future. BioFire may also add bullet points to its core sales and marketing presentations citing the modeling study, and the study could end up in the hands of BioFire's sales force in the form of reprints, he said.

Other recent studies using BioFire's syndromic respiratory panel in pediatric populations have shown benefits in terms of length of stay and reduction in antibiotic usage as well as faster turnaround times.

However, a unique challenge of the pediatric population may be the incidence of viral and bacterial carriage among kids.

For example, kids under 3 are more likely to carry Clostridium difficile — one of the pathogens identified in the FilmArray gastrointestinal panel — but not have symptoms of active infection. While detecting carriage may activate costly hospital quarantine procedures, it could also potentially benefit the patient if it informed antibiotics prescribing, as antibiotics are known to cause microbiome imbalance and C. diff overgrowth.

BioFire's respiratory and gastrointestinal panels are cleared by the US Food and Drug Administration for use in patients of all ages suspected of having infections, Stevenson noted.

"Right now we do not offer a pediatric versus an adult panel," he said. "We just have one panel, because we feel like the syndromic approach … asks the question 'What's going on in this patient's GI tract from an infectious disease stand point?' and C. diff may or may not be part of the answer."

Stevenson also provided an update on BioFire's panel pipeline, noting that its meningitis/encephalitis assay is in the FDA's hands now and will likely launch this summer.

The firm is also on track for a 2017 launch of a "pneumonia panel" that will be the most ambitious yet. It will include "a lot of bacterial and antibiotic resistance targets, a good number of viral targets, and some fungal targets as well, and it will be our first panel that has quantitative results," Stevenson said.

Charles River Laboratories International, Inc. announced today that it has entered into a definitive agreement to acquire Celsis International Ltd. for $212 million in cash, subject to customary closing adjustments. Celsis is a leading provider of rapid bacterial detection systems for quality control testing in the biopharmaceutical and consumer products industries. Celsis’s systems are principally used for product-release testing to help ensure the safe manufacture of drugs and consumer products. The acquisition complements Charles River Endotoxin and Microbial Detection’s (EMD) position as a leading provider of rapid endotoxin testing and bacterial identification for biopharmaceutical manufacturing, and creates the most comprehensive solution for rapid quality control testing of biopharmaceutical and consumer products.

The addition of Celsis, with its Advance II™, Accel™, and Innovate™ systems for non-sterile applications, will complement EMD’s recently introduced PTS-Micro™, a rapid bacterial (bioburden) detection system for sterile biopharmaceutical applications. The comprehensive EMD and Celsis portfolio is expected to drive increased adoption of EMD’s quality control testing solutions across both sterile and non-sterile applications, as clients seek a single provider for their rapid product-release testing needs.

James C. Foster, Chairman, President, and Chief Executive Officer of Charles River Laboratories, commented, “The acquisition of Celsis combines leading providers of endotoxin and bacterial testing and bacterial identification to establish EMD as a comprehensive provider of rapid quality control testing solutions across multiple industries. The acquisition nearly doubles the market opportunity for EMD testing products and services, offering access to the consumer products market in addition to our core biopharmaceutical market. Celsis’s robust financial profile enhances the EMD business, which is our highest-growth and one of our highest-margin businesses. We expect Celsis to be immediately accretive to non-GAAP earnings per share in 2015, with a more significant contribution in 2016 and beyond.”

Friday, July 3, 2015

A test kit that diagnoses Ebola rapidly using just a finger prick of blood could save lives in the ongoing epidemic in West Africa. But researchers are perplexed as to why the diagnostic kit has not been deployed in the field, despite both the World Health Organization (WHO) and the US Food and Drug Administration approving it four months ago for emergency use.

“A lot of people are frustrated,” says Nira Pollock, an infectious-diseases researcher at the Boston Children’s Hospital in Massachusetts, and the senior author of an independent field-validation study of the test kit, published on 26 June1 in The Lancet. The results were presented to the WHO and health ministries in affected countries in March. “Many groups on the ground would like to deploy the test, but can’t, because national regulators haven’t approved it.”

The test, developed by the medical-diagnostics company Corgenix, of Broomfield, Colorado, detects Ebola specific proteins that are in the blood. Like a pregnancy test kit, it shows a positive result by coloured bands; it is simple enough to use that healthcare workers with minimal training could deploy it in remote villages and get a fast result. Existing ‘gold-standard’ methods of diagnosis require blood samples to be drawn and transported to sophisticated central laboratories for processing — which can result in long delays. The Corgenix test could not replace lab confirmation, but it would allow workers to identify infected people and isolate them faster, greatly reducing the spread of disease, says Nahid Bhadelia, an infectious-diseases physician at the Boston University School of Medicine and the Boston Medical Center in Massachusetts.

At the peak of the epidemic, delays and bottlenecks in testing at centralized labs meant that people who were infectious were going undetected for around 5–6 days before being diagnosed, which had devastating effects, says Bhadelia, who is about to return to Sierra Leone for her fourth stint working in an Ebola treatment centre.

Sensitive test

One concern was that the test might fail to detect the virus in some cases of Ebola. But the independent field-validation1 (in Sierra Leone) shows that the kit was as sensitive at catching cases as the gold-standard comparison — a real-time polymerase chain reaction (RT-PCR) test that amplifies and detects genetic sequences that are specific to Ebola in blood and other bodily fluids.

Robert Garry, a virologist at Tulane University in New Orleans, Louisiana, who helped to develop the Corgenix test, blames the delays on what he says are overly restrictive and ambiguous sets of guidelines released by WHO in March and June. They state, for example, that the rapid “antigen-detection” tests “have no role in the routine management of Ebola in settings where PCR (molecular) testing is available; however, they may have utility in settings without laboratory infrastructure if their benefits and limitations are understood”.

Many people have misunderstood that to mean “if you have PCR, you don’t need the test”, Garry says. But the Ebola epidemic has shown that use of a centralized testing model alone is badly flawed, he says, and that additional decentralized testing is needed in the places where outbreaks occur to slash delays in identifying those infected. “We need to rethink the conventional wisdom about PCR being the only answer,” he adds.

False positives

But Mark Perkins, chief scientific officer of the non-profit organization Foundation for Innovative New Diagnostics (FIND) in Geneva, Switzerland, defends the thrust of the WHO's guidelines, which FIND contributed to developing. The concern with the rapid-antigen tests, he says, is that they report too many false positives. In the independent validation, 6 of 77 people who tested negative for Ebola using PCR were declared positive by the Corgenix kit. “To roll out an assay with so many false positives is unthinkable,” Perkins says.

That is less of a problem in this instance, argues Bhadelia, as researchers are proposing to use the test only to speed up the identification of those who are positive for Ebola, before confirming the results in the lab a few days later.

Moreover, the independent validation has raised questions over the quality of some of the PCR testing that was done during the epidemic. In the study, researchers used a routinely employed, partly-automated PCR kit made by altona Diagnostics, of Hamburg, Germany. But some of the samples that the altona test scored as negative — and which the Corgenix kit scored as positive — were later confirmed to be positive using a reference-standard test kit. That is a concern, says Bhadelia, as false negatives by the altona test might have allowed people with Ebola to return to the community while they were still infectious. It also suggests the possibility that the Corgenix kit might not be as prone to false positives as Perkins suggests — though further research would be needed to clarify the kit's performance.

Field research

Perkins says that although a rapid test would have helped to triage people at the peak of the Ebola epidemic last year, when PCR labs were overwhelmed, the drastic fall in the number of cases since then has meant that labs can now easily handle the load. Rapid tests are now of “reduced relevance”, he says.

But researchers say that this misses the point. Although most Ebola cases are currently close to urban PCR labs, sporadic outbreaks still occur in rural areas. Should Ebola flare up again, it would be beneficial to have gained experience using the kits in the field, says Pollock. Deploying the kits would also provide much more data about their real-world performance, useful both for this and further outbreaks, says Mara Jana Broadhurst, a co-author of The Lancet study. who works on Ebola diagnostics for Partners in Health, a global-health organization in Boston, Massachusetts.

Ebola in West Africa clearly exposed the "logistical constraints" of using PCR alone and its weaknesses as a frontline tool in a serious epidemic, says Broadhurst. "We need to consider all options.”

Regeneron Pharmaceuticals, Inc. announced that the Proceedings of the National Academy of Sciences (PNAS) has published a paper demonstrating that the Company's proprietary VelociGene® and VelocImmune® technologies enabled the rapid identification and preclinical validation of potential candidates that could be developed for the prevention and treatment of Middle East Respiratory Syndrome (MERS). This involved the development of fully human antibodies specific to the MERS virus, as well as the creation of a genetically humanized model to test and validate these antibodies. In addition to MERS, Regeneron's proprietary antibody rapid response platform has been used to generate antibodies for Ebola virus and has the potential to address other emerging infectious diseases.

"Regeneron is committed to being part of the solution in responding to emerging epidemics, and we hope to collaborate with governments and other organizations in an effort to make our antibodies for MERS and Ebola available," said Neil Stahl, Ph.D., Executive Vice President of Research and Development at Regeneron. "Fully human antibodies hold incredible promise for treating infectious disease; however, earlier generation technologies were not optimal for rapid enough development so as to address emerging outbreaks. Our Veloci-technologies identify and produce validated fully human antibodies, already in manufacturing-ready cell lines, within months as compared to years using other methods. This holds the potential to offer a rapid response solution for emerging infectious diseases, which are an increasing threat in our interconnected world."

Regeneron is currently collaborating with the Biomedical Advanced Research and Development Authority (BARDA) of the U.S. Department of Health and Human Services (HHS) on further evaluation of both the MERS and Ebola antibodies.

Currently there are no approved medicines or vaccines to treat or prevent MERS, which causes severe respiratory tract infections and is associated with high death rates. In addition to an ongoing outbreak in South Korea, cases of MERS have been reported in the Middle East, Europe, the U.S., Africa and other countries in Asia.

Details on Study Results and the Regeneron Rapid Response Platform

Regeneron researchers utilized VelocImmune®, a platform that enables the rapid generation of fully human monoclonal antibodies, to create a panel of antibodies that block interaction between the MERS coronavirus (MERS-CoV) Spike protein and its receptor, DPP4, thus preventing virus cell entry. In parallel, VelociGene®, a large-scale mouse genomic modification platform, was used to develop a genetically humanized mouse model for the MERS infection. Although mice are not susceptible to MERS, which has stymied many research efforts, VelociGene® enabled the creation of a robust mouse model of disease. In collaboration with the University of Maryland School of Medicine, which provided access to an infectious MERS-CoV clone and conducted testing with live virus, the potential treatments were evaluated in this novel small animal model.

Laser technology to diagnose malaria — one of the world’s deadliest diseases — succeeded in early trials.

Rice University researchers tested an experimental laser device that can diagnose malaria in a few seconds. The ultra-rapid diagnostic may offer substantial cost savings in global efforts to combat malaria .

The study has been published in the July issue of Emerging Infectious Diseases.

The laser was able to correctly identify which person, among the total group of six, who were infected with Plasmodium falciparum, a parasite that causes malaria. Further, the device was able to detect the parasite in dead mosquitoes, according to a New Scientist report.

The laser works by pulsing energy into a vein of a person who is suspected to have malaria. The test can be done on the wrist or earlobe.

The laser’s wavelength is harmless to human tissue, but is absorbed by waste crystals produced by the malaria parasite as it feeds on human blood.

The waste crystals absorb the energy, warm the surrounding blood plasma, making it bubble. An oscilloscope then detects malaria by sensing the bubbles as they are popping.

The diagnosis process only takes 20 seconds.

“It’s the first true non-invasive diagnostic,” says Dmitri Lapotko of Rice University in Houston, Texas, whose team tested the device, which was able to identify which person had malaria in a group of six. The device also was able to identify the malaria parasite in dead mosquitos.

Malaria is one of the leading causes of death in the world, killing 584,000 people in 2013. The World Health Organization (WHO) reports that progress has been made. In the last decade, effective strategies of vector control, rapid diagnostics, and combination therapies have reduced malaria mortality by 47% worldwide and 58% in Africa.

SEARCH FOR COST-EFFECTIVE DIAGNOSTICS

While existing tests for malaria are already quick, taking 15 to 20 minutes, they could be simpler and less expensive. Current rapid diagnostics require blood to be taken and tested by trained health workers.

Blood has to be taken, the test has to be conducted by trained personnel to get reliable results, and extra chemical reagents must be used.

Chemical reagents used in diagnostic tests come at a significant cost. The Foundation for Innovative New Diagnostics (FIND), a nonprofit established by the WHO, estimates that reagents cost $100 million USD per year.

The experimental diagnostic is non-invasive would simplify diagnosis, not needed reagents or blood handling procedures. FIND’s chief scientific officer Mark Perkins said, “The pursuit of technologies that avoid these pitfalls, especially when as innovative as this one, is welcome,” according to the New Scientist.

Lapotko estimates that a single testing unit would cost around $15,000, which could test 200,000 people. The per-person cost could be brought down from 50 cents to 8 cents per person.

THE ROAD AHEAD: FIELD TRIALS

The team is preparing for trials in Africa and to see how the device fares in real-world field conditions. It is not yet known whether the diagnostic will work in people with lower levels of parasites in their blood.

Further, the probe is less reliable with people with darker skin, a major problem because children in African account for the major of malaria deaths. Lapotko’s team is confident that they will be able to fix this pitfall with a wavelength adjustment.

Accelerate Diagnostics, Inc., announced its declaration of conformity to the European In Vitro Diagnostic Directive 98/79 EC and CE Mark of the Accelerate ID/AST System and ID/AST Blood Culture Assay for in vitro diagnostic use. The Accelerate ID/AST System is a diagnostic platform providing rapid identification and antimicrobial susceptibility testing of serious infections.

In addition, the company initiated enrollment at 8 clinical trial sites for its preclinical study. Upon successful completion the FDA registration trial will begin. Given the aforementioned progress, the company maintains its expectation that it will launch an FDA cleared product in the United States in the first half of 2016.

As part of their effort to battle the ongoing Ebola outbreak in Sierra Leone, researchers from Partners in Health and Public Health England (PHE), as well as their collaborators in Sierra Leone's Ministry of Health and Sanitation and Geneva's Foundation for New Innovative Diagnostics, recently performed a field validation of the Corgenix ReEBOV Antigen Rapid Test kit.

Given that standard testing at the PHE field lab in Port Loko throughout the course of the outbreak in Sierra Leone had involved the use of Altona Diagnostics' RealStar Filovirus Screen RT-PCR kit, the team used that kit as a benchmark to validate the results of the Corgenix test.

They found that the Corgenix test had 100 percent sensitivity and 92 percent specificity in detecting Ebola virus from patient blood samples. However, the team also found that the Altona assay itself performed less than optimally in the PHE lab.

"Surprisingly, the findings also revealed that the standard Altona RT-PCR test, under the conditions deployed in the field, was itself an imperfect reference standard," the authors wrote in a press release. "The Altona RT-PCR assay failed to detect a small number of EVD cases that tested positive by both RDT and by an alternative RT-PCR test, all with relatively low amounts of virus."

The results were unexpected, Boston Children's Hospital's Nira Pollock, the study's senior author, told GenomeWeb. The Altona test was not as sensitive as the team had expected, especially compared to the alternative assay, previously published by a team led by the US Army Medical Research Institute of Infectious Diseases (USAMRIID).

"We didn't expect the Altona [assay] to have this sensitivity limitation in the low viral load sample set. And we only figured that out because we did discordant analysis when we saw that some samples were rapid test-positive and Altona-negative and wanted to figure out whether these were false-positive rapid tests or false-negative Altona," Pollock said.

However, she added, the problematic results can be blamed — at least partially — on the way the test was deployed in the PHE lab, including the platforms that were selected for nucleic acid extraction and amplification, like the Cepheid SmartCycler system. "That may have led the Altona to not perform as well as it could have had it been deployed differently," Pollock said.

Indeed, said Altona's Stephan Oelschlaeger, who develops kits like the RealStar FiloVirus for molecular diagnosis of tropical and emerging diseases , the problem is that the PHE lab used the Altona assay differently than the company recommends. The SmartCycler is not on the list of systems that would produce the most accurate result from the RealStar kit. In fact, Oelschlaeger added, the difference in sensitivity when the kit is used with approved cyclers than when it's used with the SmartCycler is significant. "I am confident that it works very well on the cycler platforms that we specify in our manual," he said.

Pollock seemed to concur. "The Altona package insert specifies certain extraction conditions and certain amplification platforms that it was validated with during the approval process and the PHE lab chose to use some alternative methods," she says. "So it's possible that if the lab had used exactly what was in the package insert, it would have performed better." There's research underway to determine if that's the case, she added.

Currently, Oelschlaeger says, Altona doesn't plan to make the RealStar kit compatible with the SmartCycler system. Labs that only come equipped with SmartCyclers have other options, like the USAMRIID assay. Markus Hess, Altona's head of R&D, said getting the RealStar kit to work on the SmartCycler would not be a matter of making a simple adjustment. "We have assays specifically for the SmartCycler, but it's a different concept," he adds.

But it's important to note that field labs may not all be equipped to use certain assays the way they're recommended, according to Pollock. "Different labs make different choices," to reduce turnaround times or account for sample sizes, she says. "In this case, the PHE lab used smaller volumes of samples for nucleic acid extraction … and that may have decreased the sensitivity. These choices were made for a reason, to try to improve the overall result package in terms of accuracy, speed, and reliability. They were made deliberately to try to optimize testing in an outbreak scenario."

However, she added, the study shouldn't be taken as an indication that there's something generally wrong with PCR-based tests — which are considered the gold standard. What it does is highlight some complications with molecular-based testing in conditions like the middle of an infectious disease outbreak, and shows that there's a need for better point-of-care tests to begin diagnosis.

"It's important to recognize that molecular methods aren't perfect," Pollock says. "They have problems too, particularly in a complicated, dirty laboratory setting in the field. You can have contamination or inhibition. And when you send samples anywhere to another place to have them tested, there are all sorts of issues — not just the transport times, but also the paperwork, samples coming with wrong patient identifiers, results getting reported to the wrong person or the wrong place. And then even if you get results in the lab, you still have to get results out to the clinicians — there could be internet problems or issues with transport of the paper result file." All of these circumstances must be taken into account when considering what kind of test to use to diagnose patients.

"Everyone is always focused on test accuracy, but there's so much more than that," she adds.

For its part, Corgenix is very pleased with the study's results — the company's infectious diseases consultant, Doug Simpson, said he was surprised to see the 100 percent sensitivity. Corgenix will continue to make its rapid diagnostic kit available for any testing being done in West Africa, he adds. The company is also continually improving the test's antibodies so as to be ready if another Ebola outbreak occurs. "We need bigger, better, faster tests so that it doesn't get out of hand like it did last year," Simpson said.

Outbreak preparedness is one goal of the RealStar assay, Oelschlaeger said. In that context, it would be more likely that the kit would be used in a lab that has the recommended cyclers, which would help the assay perform at optimal levels.

The big question now, Pollock said, is how to use all of these tests to get the most accurate diagnostic information. Indeed, she added, what should be done in a large setting is to implement the Corgenix test in conjunction with confirmatory PCR. That way, researchers and doctors can learn the best testing methods to use in different stages of a potential outbreak, and throughout the years that follow. "We can't even speculate how long clinicians in West Africa are going to [need] to test for Ebola if someone presents with compatible symptoms," she said. "Maybe it's forever."

The AOAC Research Institute (AOAC-RI) has granted a method extension for the DuPont™ BAX® System real-time polymerase chain reaction (PCR) assays for Listeria to include validation of a new proprietary enrichment media. This method can return Listeria results in just 22 hours for some matrices--one of the fastest time-to-results available for the pathogen in the food safety industry today.

The AOAC-RI method allows customers to perform a single-stage, 20-26 hour enrichment (sample-dependent) in FoodChek™ Actero™ Listeria Enrichment Media, followed by rapid real-time processing with the DuPont™ BAX® System. The extension includes detection of both Listeria species (AOAC-RI Performance Tested MethodSM #081401) and L. monocytogenes (AOAC-RI Performance Tested MethodSM #121402) and has been tested in frankfurters, shrimp, smoked salmon, spinach and queso fresco, as well as on stainless steel, plastic and sealed concrete surfaces.

“The biggest advantage of this combined testing method is the reduced enrichment time, without the need for a secondary enrichment protocol,” says Morgan Wallace, DuPont Nutrition & Health senior microbiologist and validations leader for diagnostics. “Listeria is a slow-growing pathogen, so any steps that allow for faster detection are important to the food safety industry and help provide cost-saving benefits to customers.”

“Enriching food samples in Actero™ Listeria Enrichment media allows laboratories to use about 33 percent less media per sample than other methods,” says FoodChek™ chairman, president and CEO William J. Hogan. “This not only results in less waste disposal during the testing process, but also helps improve the financial bottom line for food companies and commercial testing laboratories alike.”

The DuPont™ BAX® System provides advanced, automated testing for foodborne pathogens, spoilage organisms and other microbes in raw ingredients, finished products and manufacturing environments. The system breaks down samples at the genetic level using the power of the PCR, then uses target-specific primers to automatically detect the presence or absence of the target bacterial DNA and provide clear yes-or-no results. With certifications and regulatory approvals around the world, the BAX® System is recognized as one of the most advanced pathogen testing systems available to the food industry today.